Single-cell mRNA transfection studies: Delivery, kinetics and statistics by numbers

被引:88
作者
Leonhardt, Carolin [1 ,2 ]
Schwake, Gerlinde [1 ,2 ]
Stoegbauer, Tobias R. [1 ,2 ]
Rappl, Susanne [1 ,2 ]
Kuhr, Jan-Timm [3 ]
Ligon, Thomas S. [1 ,2 ]
Raedler, Joachim O. [1 ,2 ]
机构
[1] Univ Munich, Fac Phys, Munich, Germany
[2] Univ Munich, Ctr NanoSci CeNS, Munich, Germany
[3] Tech Univ Berlin, Inst Theoret Phys, Berlin, Germany
关键词
mRNA transfection; Non-viral gene delivery; Expression kinetics; Single-cell studies; Pharmacokinetics; SYNTHETIC GENE DELIVERY; LIPOSOME-DNA COMPLEXES; CAP-ANALOGS; DENDRITIC CELLS; TRANSCRIPTION; EFFICIENCY; EXPRESSION; STABILITY; POLYPLEX; EFFICACY;
D O I
10.1016/j.nano.2013.11.008
中图分类号
TB3 [工程材料学];
学科分类号
0805 ; 080502 ;
摘要
In artificial gene delivery, messenger RNA (mRNA) is an attractive alternative to plasmid DNA (pDNA) since it does not require transfer into the cell nucleus. Here we show that, unlike for pDNA transfection, the delivery statistics and dynamics of mRNA-mediated expression are generic and predictable in terms of mathematical modeling. We measured the single-cell expression time-courses and levels of enhanced green fluorescent protein (eGFP) using time-lapse microscopy and flow cytometry (FC). The single-cell analysis provides direct access to the distribution of onset times, life times and expression rates of mRNA and eGFP. We introduce a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby the dose-response relation. Our results establish a statistical framework for mRNA transfection and as such should advance the development of RNA carriers and small interfering/micro RNA-based drugs. From the Clinical Editor: This team of authors established a statistical framework for mRNA transfection by using a two-step stochastic delivery model that reproduces the number distribution of successfully delivered and translated mRNA molecules and thereby their dose-response relation. This study establishes a nice connection between theory and experimental planning and will aid the cellular delivery of mRNA molecules. (C) 2014 The Authors. Published by Elsevier Inc. All rights reserved.
引用
收藏
页码:679 / 688
页数:10
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